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ACCURIS INSTRUMENTS PR6000 Terminator v3.1 Cycle Sequencing Kit
លក្ខណៈបច្ចេកទេស
- Component: Reaction Mix
- Limitations of Use: For research purposes only, not for therapeutic or diagnostic use
ការណែនាំអំពីការប្រើប្រាស់ផលិតផល
- Accuris SmartDye Terminator v3.1 Cycle Sequencing mix is a direct replacement for Applied Biosystem BigDye v3.1.
- It offers a faster reaction cycle and a higher success rate with difficult templates.
- No changes in protocol are needed, and no new software or calibration is required.
- This protocol covers the reaction setup and cycle sequencing parameters.
- For detailed instructions on template preparation, purity, and sequencing reaction purification, refer to your instrument manual.
- Research shows longer annealing time helps with GT richtemplates. [2] 3-minute extension allows reading length of 1000bp – 1200bp.
- Catalog number PR6000-1K includes 4x2ml of dye, 12ml 5X buffer
- Catalog number PR6000-5K includes 2x20ml of dye, 56ml 5X buffer
- Catalog number PR6000-25K includes 10x20ml of dye, 466ml 5X buffer
Accuris™ SmartDye™ Terminator v3.1 Cycle Sequencing Mix
ការពិពណ៌នា
- Accuris SmartDye Terminator v3.1 Cycle Sequencing mix is a direct replacement for Applied BioSystem™ BigDye™ v3.1. No changes in protocol are needed and no new software or mobility shift file needs to be installed in the system.
- There is no need for new calibration or validation.
SmartDye Advantages
- Faster reaction cycle. A 3-minute extension time is utilized instead of the 4 mintues required by BigDye. This can be reduced to 2 minutes for many templates.
- Higher dilution factor. As little as 0.15μl of mix can be used in a 5μl reaction volume.
- Higher success rate with difficult templates.
ការផ្ទុក
- Upon receipt, immediately store the SmartDye Mix at-20°C to -80°C. The 5X sequencing buffer should be stored at 4°C to -20ºC. Avoid excessive freeze/thaw cycles.
- Protect dye from light. If needed, reagents may be aliquotted into smaller amounts.
- Before use, thaw the frozen stocks on ice, mix thoroughly and centrifuge briefly to bring all the mix to the bottom of the tube.
ដែនកំណត់នៃការប្រើប្រាស់
- សម្រាប់គោលបំណងស្រាវជ្រាវតែប៉ុណ្ណោះ។ មិនមានបំណងសម្រាប់ការប្រើប្រាស់ព្យាបាល ឬរោគវិនិច្ឆ័យទេ។
- Accuris មិនទទួលខុសត្រូវចំពោះការខូចខាតជាលទ្ធផល ឬដោយចៃដន្យ មិនថាដោយផ្ទាល់ ឬដោយប្រយោល ដែលបណ្តាលមកពីការប្រើប្រាស់ផលិតផលនេះទេ។
- Accuris ធានានូវដំណើរការនៃផលិតផលនេះ ដូចដែលបានពិពណ៌នានៅពេលប្រើដោយអនុលោមតាមការណែនាំទាំងនេះ។ វាជាទំនួលខុសត្រូវរបស់អ្នកទិញក្នុងការកំណត់ភាពសមស្របនៃផលិតផលនេះសម្រាប់កម្មវិធីជាក់លាក់របស់ពួកគេ។
គោលការណ៍ណែនាំទូទៅ
This protocol covers the reaction setup and cycle sequencing parameters. For detailed instructions on how to prepare the templates, template concentrations, purity and sequencing reaction purification, etc, please keep your current protocol unchanged or refer to your instrument manual.
- Reactions can be set up in 0.2ml single PCR tubes or 96 and 384 well plates according to the formula below.
- We recommend 5μl or 10μl total reaction volumes. Please read the footnotes carefully.
| សមាសភាគ | 5µl ប្រតិកម្ម | 10µl ប្រតិកម្ម |
| Template [1],[2] | 2µl | 2µl |
| Primer (5µM) | 1µl | 1µl |
| 5X Seq Buffer [3],[5] | 0.85µl | 1.7µl |
| SmartDye Mix [4],[5] | 0.15µl | 0.3µl |
| ទឹក។ | 1µl | 5µl |
| បរិមាណសរុប | 5µl | 10µl |
- 150-300ng/μl of dsDNA
- The concentration of the template may affect volume.
If the template volume differs, adjust the amount of water added to keep the final volume the same. - The volume of 5X buffer needed per reaction is determined by the following formula:
- A maximum of 0.5μl or 1.0μl of SmartDye Mix is recommended for 5μl and 10μl reactions, respectively. Using more may give less than favorable results.
- A ready mix cocktail can be made by mixing 100 volumes of water, 85 volumes of 5X sequencing buffer, and 15 volumes of SmartDye Mix. Use 2μl or 4μl of the ready mix for each 5μl or 10μl reaction.
Perform the cycle sequencing in a thermal cycler:
| ជំហាន | សីតុណ្ហភាព | ពេលវេលា |
| ការប្រែពណ៌ដើម | 94°C | 1 នាទី។ |
| 30 វដ្ត | 94°C | 10 វិនាទី |
| 50°C | 10 seconds [1] | |
| 60°C | 3 minutes [2] | |
| កាន់ | 4°C | កាន់ |
- Research has shown that longer annealing time facilitates sequencing through GT-rich templates.
- An extension for 3 minutes allows a reading length of 1000bp – 1200bp. Shorter extension times can be used for shorter templates.
ជំនួយបច្ចេកទេស
- For troubleshooting and tech support, contact us by phone at 908 769-5555 or email info@accuris-usa.com.
- When possible, please include instrument model, reaction conditions, PCR parameters, amplicon size, and any traces and melting profiles.
មាតិកាកញ្ចប់និងការបញ្ជាទិញឡើងវិញ
- Accuris SmartDye Terminator v3.1 Cycle Sequencing Kit is supplied with buffer, control and primer.
24 ប្រតិកម្ម
- Catalog number PR6000-24
- Includes 200μl of dye, 1ml 5X buffer
100 ប្រតិកម្ម
- Catalog number PR6000-100
- Includes 800μl of dye, 1ml 5X buffer
1,000 ប្រតិកម្ម
- Catalog number PR6000-1K
- Includes 4x2ml of dye, 12ml 5X buffer
5,000 ប្រតិកម្ម
- Catalog number PR6000-5K
- Includes 2x20ml of dye, 56ml 5X buffer
25,000 ប្រតិកម្ម
- Catalog number PR6000-25K
- Includes 10x20ml of dye, 466ml 5X buffer
ទំនាក់ទំនង
- PH: 908 769-5555 EM: info@accuris-usa.com
សំណួរគេសួរញឹកញាប់
សំណួរ៖ តើផលិតផលនេះអាចប្រើសម្រាប់គោលបំណងវិនិច្ឆ័យបានទេ?
A: No, this product is for research purposes only and not intended for therapeutic or diagnostic use.
សំណួរ៖ តើអ្វីជា advantage of Accuris SmartDye Terminator v3.1 over BigDye v3.1?
A: Accuris SmartDye Terminator v3.1 offers a faster reaction cycle and higher success rate with difficult templates compared to BigDye v3.1.
ឯកសារ/ធនធាន
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ACCURIS INSTRUMENTS PR6000 Terminator v3.1 Cycle Sequencing Kit [pdf] សេចក្តីណែនាំ PR6000 Terminator v3.1 Cycle Sequencing Kit, PR6000, Terminator v3.1 Cycle Sequencing Kit, v3.1 Cycle Sequencing Kit, Cycle Sequencing Kit, Sequencing Kit |